Mannoprotein Cig1 Contributes to the Immunogenicity of a Heat-Killed F-box Protein Fbp1 Cryptococcus neoformans Vaccine Model

Currently, no fungal vaccine exists for clinical use while fungal infections are responsible for over 1.5 million deaths every year. Our previous studies identified a Cryptococcus neoformans mutant strain fbp1 Δ as a potential vaccine candidate. This mutant strain contains a deletion of the F-box protein Fbp1, a key subunit of the SCF E3 ligase complex necessary for ubiquitin-mediated proteolysis. Vaccination with heat-killed fbp1 Δ (HK- fbp1) can elicit protection against C. neoformans parental strain and its sibling species C. gattii in an interferon gamma (IFN-γ) dependent Type 1 immune response. However, we have yet to decipher the immunogenic factor(s) expressed by the fbp1 Δ mutant that are responsible for the induction of the protective immune response. In this study, we have identified that capsule plays an important role in HK- fbp1 vaccine mediated protection, as acapsular HK- fbp1 cells showed diminished protection against wild type challenge. Additionally, our studies have shown that Cytokine Inducing Glycoprotein 1 (Cig1), a GPI anchored mannoprotein, is regulated by Fbp1 and contributes to the immunogenicity of HK- fbp1 . Deletion of Cig1 in the fbp1 Δ background resulted in decreased recruitment of anti-fungal effector T cells and diminished production of protective inflammatory cytokines by the host. Furthermore, loss of Cig1 in the fbp1 Δ mutant resulted in reduced protection in vaccination survival studies at lower vaccine inoculum doses compared to HK- fbp1 . In aggregate, these findings demonstrate Cig1 is an antigen contributing to the immunogenicity of HK- fbp1 that may be utilized to further optimize the HK- fbp1 fungal vaccine as a tool in the arsenal against invasive fungal infections.